JPH07500418A - Apparatus and method for aligning capillary separation tubes and detection optics - Google Patents

Abstract

PCT No. PCT/US92/08589 Sec. 371 Date Mar. 29, 1994 Sec. 102(e) Date Mar. 29, 1994 PCT Filed Oct. 8, 1992.An apparatus and method of aligning a capillary with respect to a radiation source. More particularly, the capillary is aligned with a laser beam for laser induced fluorescence detection. The scatter or transmitted light pattern of the laser beam with respect to the capillary tube is utilized to determine optimum alignment. photosensors may be implemented to detect the light pattern which represents optimum alignment. For dynamic alignment during electrophoresis, the photosensors provide feedback to a controller which controls a positioning mechanism for alignment of the capillary.

Description

[Detailed description of the invention] BACKGROUND OF THE INVENTION Apparatus and Method for Aligning Capillary Separation Tubes and Detection Optics field of invention The present invention relates to the optical detection of macromolecular components separated in a capillary separation tube, and and relating to the alignment of the capillary separation tube with the detection optics.

Description of related technology Macromolecules in biological samples can be separated into capillary tubes (ca. can be separated in a pillary column). One of the techniques One is electrophoresis of tubules. In this, the sample is placed at a height along the channel of the tubule. are separated within the channels of the tubules by applying a high potential. The sample is huge Differences in the separation medium that are inherent to the molecule and depend on the charge, size, and shape of the macromolecules As a result of their mobility, the macromolecular species are separated into zones.

Separated zones are detected by several techniques. Optical detection of multiple One of the techniques is the detection of laser-induced fluorescence. The detection technique No. 4, U.S. Pat. No. 4, to Zare et al. There is a specification of No. 675,300. Generally, light from a laser source is sent to a separate sample. passes through the cross-section of the separation channel of the capillary to generate the liquid component. Said sample Prior to, during, or immediately after separation, the components are provided with a fluorescent material to fluoresce. A highly luminescent material is added. The fluorescence can be detected with high sensitivity. high feeling For maximum resolution and maximum resolution, short tubular separation channels define a small detection volume. Focus the laser beam on a narrow spot to interrogate the cross section This is achieved by The typical size of the spot is 50 microns. For separation channels with outer diameters on the order of 50 microns or larger on the order of or smaller.

Similar to the optical detection scheme used, the highest sensitivity depends on the overall laser beam configuration, the capillary Only when the separation channel and optical elements are properly aligned can the laser-induced achieved for luminescence detection. The critical components of proper optical alignment are: This is the positioning of the detection volume of the capillary with respect to the laser beam. Because typical The output volume and the dimensions of the capillary are very small, and the interaction between the channel of the capillary and the laser beam is very small. This is because misalignment of just a few microns can significantly degrade detection performance. Ru.

Correct placement of the capillary in the laser beam to achieve perfect alignment and It is difficult to maintain this arrangement once achieved. mix this difficulty This is due to the condition that the capillary can be easily replaced and the fact that the capillary is exposed to a high electric field. and the fact that alignment shifts as a result of thermal fluctuations. including. The exact cause of tubule movement is uncertain. The electric current on the inner wall of the tubule separation tube The charge distribution within the tubular channels associated with time-dependent changes in the field is partly responsible. Some people doubt that there may be one.

Oven tube capillary (filled with gel electrolyte or similar substance) (as opposed to a capillary filled with electrolyte solution) prior to performing electrophoresis. One recent effort to match is to supply a fluorescent solution to a capillary, which can be connected to a photomultiplier tube or adjusts the alignment while monitoring the fluorescence with a photodiode. The alignment is It is considered that this was achieved when the fluorescence intensity was at its highest.

The alignment means described above are the tubes of the oven duplex, i.e., non-gel (liquid) Applicable only to capillaries filled with electrolyte only. Before it is aligned, the fluorescent solution is The gel cannot be effectively sucked out into the separation tube filled with gel, and the gel cannot be removed after alignment. This is because they cannot be completely eliminated. This means also that the oven tube It is often difficult to completely remove the fluorescent substance from the inner walls of the tubules, and Even oven tubes are undesirable, since this is not possible.

Moreover, none provide for dynamic alignment during the electrophoretic process. Thin Tube Separation Tubes become laterally misaligned under thermal fluctuations or high applied potentials. Therefore, when the alignment shifts from the optimal state during the electrophoretic operation, There's no way to know that.

Summary of the invention The present invention provides a method for observing the light pattern or patterns formed by the tubules. A device for aligning the capillary separation tube with respect to or with respect to the radiation source by Directed to the location and method. In one aspect of the invention, the alignment comprises tubular channels. A unique and reproducible scattering pattern of radiation that occurs at the interface between the wall and the contents of the tubule. i.e. by relying on the scattering pattern. Ru. In another aspect of the invention, the alignment is uniquely reproducible observed after passing through the tubules. This is accomplished by relying on a conducted beam pattern. Still another aspect of the present invention For surfaces, alignment relies on distinct shadows cast onto the surface by elements of the optical system. It is fulfilled by this. The method described here detects the presence of fluorescent materials. It is not necessary and can be applied to both oven tube tubes and gel-filled tubes. Wear. The alignment procedure requires proper alignment of the tubular channel with respect to the radiation source. It can be carried out at any stage of the process, for example during electrophoresis.

In a first aspect of the invention, radiation from a light source, for example a laser beam, Focused on separation channel. While looking through the microscope objective lens, The tube is moved laterally across the radiation source. This is the inner wall of the tubule and the contents of the tubule A distinct scattering pattern starting from the interface between the objects is perpendicular to the laser beam. This is done until a plane of diameter in the direction is observed. In a second aspect of the invention, the alignment is , determined from the far-field image of the transmitted beam pattern. Laterally through the tubules Distinct beam pattern formed by the conducted laser for optimal alignment is observed. In the third plane, the radiation scattered from the capillary is normalized and the scattering block is Used to cast a shadow on the stop piece. Optimal laser and/or tubule alignment A specific shadow corresponding to is observed.

A photosensor can be employed to detect any one of the light patterns described above. Ho The output of the sensor is applied to control the servomechanism. The servo mechanism is optimal Laser beam separation channels of tubules to create patterns corresponding to optical alignment Adjust the horizontal position relative to the frame. In other words, the photosensor achieves dynamic alignment. This provides feedback on the matching conditions to ensure that the matching is consistent with the electrophoretic operation. Continuously monitored, calibrated and maintained during production.

Brief description of the drawing FIG. 1 is a diametrical cross-sectional view of a capillary tube.

Figure 2 is a schematic diagram of a capillary electrophoresis device with laser-induced fluorescence detection. be.

FIG. 3 is a diametrical cross-sectional view of the sensing section of the capillary tube.

Figure 4 shows various positions of the laser beam with respect to the separation channel of the optical fiber capillary. FIG.

Figure 5 shows the scattering pattern observed with the microscope objective for the various positions in Figure 4. FIG.

FIG. 6 is a diagram schematically showing the dynamic alignment device of the present invention.

FIG. 7 is a schematic diagram showing the screen of the aperture plate.

Figure 8 shows a scattering image observed in the far field without using a microscope objective lens. FIG.

Figure 9 shows the conducted beam pattern observed on a diameter across a capillary tube. FIG.

10A to 10c show that the laser beam is translated across the capillary tube. FIG. 4 shows the appearance of a transmitted beam when

FIG. 11 is a schematic diagram of another embodiment of a dynamic alignment device.

FIG. 12 is a circuit diagram of one embodiment of feedback control.

FIG. 13 is a schematic diagram showing fluorescence collection and criteria using a baraboloid reflector. be.

14A to 14c show the parabola of the blocking strip at various positions of the laser beam. FIG. 3 is an image showing the shadow associated with the focal point of the id reflector.

Description of the illustrated embodiments The following description is of the best presently contemplated mode of carrying out the invention. child The following description is made for the purpose of illustrating the general principles of the invention and is not to be taken in a limiting sense. Shouldn't. The scope of the invention is best determined with reference to the appended claims. Ru.

The present invention hereinafter describes the detection of laser-induced fluorescence in capillary electrophoresis. Although described with reference to However, it can also be applied to the alignment of the tubular channel and the radiation source in other situations. That is understood.

The capillary tube 10 is shown in diametrical section in FIG.

The tube is a capillary on the order of 5-500μ, typically smaller than 200μ. It defines a separation channel 12 of cylindrical dimensions. Cylindrical wall of capillary tube 10 14 can be made of organic materials such as glass, fused silica or Teflon. Ru. To strengthen the capillary tube, a polyimide coating 15 is applied to its outer surface. It is glued.

Capillary tube 10 is generally flexible. i.e. bent into a smooth curve can be done.

FIG. 2 shows a capillary electrophoresis device 16, more particularly in which detection is laser induced. 1 shows a schematic arrangement of an electrophoresis device achieved by fluorescence. capillary chew The two ends of the bullion are immersed in the electrolyte 18 contained in the reservoir 20.21. It is. High voltage source 2 capable of applying high electric fields (typically l-30KV) 4 electrically connects the electrolyte 18 in the reservoir 20.21 using the electrode 22.23. It is connected. The separation channel may contain an electrolytic solution, gel electrolyte or other suitable It is filled with a separating support medium, which can be a conductive medium. Before electrophoresis Thus, the sample to be electrophoretically separated is located at one end of the separation channel 12. It is injected into the body. This can be done by any of the conventional techniques.

With the two ends of the capillary tube 10 immersed in the electrolytic solution 18, its components are separated. The high voltage supply 24 is turned on to cause electrophoresis of the sample resulting in It will be done.

In order to be able to detect the fluorescence of the separated components of the sample, The fluorescent substance is loaded with a fluorescent substance such as fluorescein. Laser-induced For high light detection, a laser 26 is used as an excitation light source. The laser 26 , for example, an air-cooled argon ion laser or a helium-neon laser. Helium-cadmium laser with output power in the range of 0.2-100mW . A 5ou optical fiber 28 is lasered into the detection section along the capillary tube 1o. Can be used to direct output. Alternatively, the lens focuses the beam directly onto the capillary. (not shown). This section describes the protective coating of polyimide. The ring 15 allows the laser light to reach the separation channel 12 through the capillary wall 14. (See Figure 3). Instead of using a laser source, other radiation sources can be used. It can be used to excite the fluorescence of the fluorescent tag of the sample component.

The laser beam is directed at 90'' to the axis of the capillary tube 10. When the sample component passes through the laser beam, the sample component should fluoresce. The photomultiplier tube 3o uses only the optical fiber 28 to detect fluorescence. (can also be placed at right angles to the capillary tube 1o. The time of detected fluorescence is Separated components can be separated by selecting different fluorescent tags for pulling them apart. components are confirmed.

Besides fluorescence, the laser beam causes light scattering in the capillary tube. vinegar. As used herein, the term scattering also refers to the reflection, refraction, diffraction and and other mechanisms in which light encounters materials in its path or changes in refractive index. This includes a mechanism that causes deviation from the incident direction. All the scattered light is , but not necessarily with the same strength; the strongest component is in the direction of the capillary tube 1 It lies in a plane perpendicular to the zero axis. In particular, the inner and outer interfaces at the walls of said tubules The interface scatters the laser beam in all directions in a plane perpendicular to the capillary tube. cause 2. Referring to FIG. 3, the diameter of the detection section of the capillary tube 10 The upper section is shown in detail. Interface between the outer surface of the tubule and the atmosphere 34.35 changes in the refractive index between the inner surface of the tubule and the contents within the tubule channel 12. Due to the change in the refractive index at the interfaces 36 and 37, the laser beam Scattered at the interface. The laser beam has a diameter of the capillary tube 10, i.e. , when the light is aligned in a diametrical plane from the four positions 34-37 identified in FIG. It can be seen that scattering is easy (as can be seen). This is one fundamental aspect of the present invention. It is a premise.

Previous researchers intentionally avoided the effects of light scattering on fluorescence detection. figure As shown in Fig. 3, the unique and reproducible scattering originating from the walls of the tubules was advantageously used. There is no one. Because the present invention exhibits optimal matching of the radiation beam and the separation channel, Using the unique reproducible scattering pattern. To assist in further explanation of the invention. For this purpose, a coordinate system is defined in Figure 3. The axis of the capillary tube is along the X axis The laser beam is along the x-axis perpendicular to the capillary tube and the y-axis are perpendicular to the X-axis and the X-axis.

Using the microscope objective 62 (Figure 4) along the y-axis focused on the diametrical plane When the scattering pattern observed in the y-axis at different stages of alignment, i.e. The beam is located at various positions A-H along the arrow 29 in the y-axis with respect to the tubule. The resulting scattering pattern is shown in FIG. Positions A-E are shown in the cross-sectional view of Figure 4. It is shown. Each scattering band shown in Figure 5 is associated with the (inner or outer) surface of the tubule and ( starting from the interface between adjacent media (air or separation support media). rank In position A, the laser beam does not impinge on the capillary tube IO. At position B, the recorder The laser beam is hitting the capillary tube 10, but it is aligned with the center of the capillary tube 10. It doesn't match. A bright band 50 of light appears above the left side of the capillary tube 10 (FIG. 4). Observed on the outer surface 40, a less bright band 51 is observed above the left side of the tubule wall 14. Observed on the inner surface 41. Between the inner surface 42 and the outer surface 43 of the lower wall of the tubule Because of the angle to the laser beam, the scattering bands 52,53 in these planes are invisible. There is nothing to stand on.

At position C, the laser beam is aligned with the center (diametrical plane) of the capillary tube. . That is, it is in the optimal alignment position for laser-induced fluorescence. This much At the position, two distinct bright bands 54,55 appear on the inner surface 44 and above the wall of the tubule. and the lower inner surface 45. Two separate bright bands 56.57 It is observed on the upper lateral surface 46 and the lower lateral surface 47 of the tubule wall. microscope Since the objective lens focuses on a plane on the diameter of the capillary tube, band 5 4.55 is brighter than the band observed when the tubule tubes are not in optimal alignment. , appears clearly.

In position, the scattering bands 58, 59 are located on the inner and outer surfaces 48 and 59 below the walls of the tubule. At 49, a pattern opposite to that observed at location B is observed. at position E , the laser beam is outside the capillary tube 10 as in position A.

The scattering patterns shown in Figures 3 and 5 focus on a diametrical cross-section of the capillary tube. It is specifically mentioned that the image is an image observed with a matched objective lens. scattering When the light is seen in the far field without an objective lens, the image of scattering is perpendicular to the tubular tube. It will be appreciated that this appears as a piece 90 (FIG. 8).

Referring to FIG. 6, which schematically shows one embodiment of an alignment device, a The optical fiber 28 is supported by a micro-positioning mechanism 60. My The cross-positioning mechanism 60 allows adjustment of the optical fiber 28 along the X and Y axes. do. The objective lens 62 of the microscope is illuminated by a laser beam through the optical fiber 28. Focus on the diametrical plane of the detection section of the capillary tube 10 being irradiated. (The capillary tube 10 is on the X axis and perpendicular to the plane of the figure). detection section The image of the image is collected on the screen defined by plate 64. plate 6 4 has an opening 65 as shown in FIG. Medial surface above and below the tubule wall The image of the scattering band 54,55 starting from the inner surface of the aperture 65 is 67 and the lower edge 68. Aperture 65 separates the capillary from the separation channel. Most of the fluorescent light is allowed to pass through to the photomultiplier tube 70, while the The scattering bands 56.57 as well as the scattering bands 54.55 starting from the outer surface It also prevents The alignment of the optical fiber 28 to the capillary 10 is such that the scattering band 54.5 5 sharply and with approximately equal strength to the edges of the rectangular aperture (or to the calibration). It is optimal when it appears (with a dependent specific strength ratio). Therefore, laser induced Usually considered as an unwanted source of background signal within the fluorescent light Source scattering is added to optimize optical alignment. The matching method described above The method uses a well-defined scattering pattern of radiation to optimize optical matching. at the same time reducing scattering from the fluorescent signal.

alignment is the result of thermal fluctuations, or the high electric fields applied to electrophores the sample; It was observed that this can vary. The latter, probably.

Electrical tension within the capillary tube lO that causes the capillary tube to deflect, or due to static electrical interaction between the fiber and the optical fiber 28. Therefore, electrophoresis During the movement, it is necessary to dynamically realign the optical fiber 28 and the capillary tube 10.

Dynamic realignment is performed using a rectangle in which the scattered bands are expected to be brought together at the best alignment. By placing a photosensor 76.78 near each edge 67.68 of the aperture 65, can be made possible. The signals from the photosensors 76 and 78 are as follows.

It is input to the controller 80. The controller 80 moves the optical fiber 28 in the y-axis direction. The movement of a servo motor, ie, a pulse motor 82, for translational movement in the direction is controlled.

The controller causes the motor to search for an optimal alignment position. This consists of two Photosensor 76.78 detects scattering band 54.55 and the band has the same intensity. achieved when it is detected that the It will be done. The circuitry and logic of controller 80 provides the required matching mechanism to be achieved. This will be obvious to those skilled in the art who are not given the ability to do so. Therefore, I will not elaborate here. stomach. Prepare to calibrate or balance the response of two photosensors due to manufacturing tolerances It is understood that this must be done.

The most important alignment of optics for detection purposes is the laser beam, i.e. the optical fiber 28 and the capillary tube 10. Microscope objective lens 62 The focusing of the capillary tube 1o on the laser beam and the capillary tube 10 is The consistency between them is not important. Fluorescence detection systems are By aligning the tubular tube 10, the angle of the tubular tube 1° with respect to the objective lens 62 of the microscope is determined. Relatively insensitive to focusing.

Signals from the two photosensors 76 and 78 are transmitted through the capillary tube 10 and the optical fiber 28. Provide feedback to automatically adjust alignment. Objective lens of IJ mirror said aperture to provide feedback to automatically adjust the focusing of the lens. Additional photosensors can be added along the edge.

The photosensor and aperture combination plays a dual role here. The opening block The rate is the scattering band that passes fluorescent light from the contents of the capillary to the photomultiplier tube 70. It acts as a spatial filter that substantially physically blocks the waves. The opening brake The photosensor provided at the top connects the laser beam to the separation channel of the capillary. provides a means of feedback that can be used to automatically align the This method According to is especially mentioned. In fact, alignment is maintained continuously during electrophoresis. why If so, matching is based on the appropriate scattering pattern of the source rather than the sample fluorescence. The alignment of tubules filled with gel or other stabilizers is based on imaging. It is possible to optimize.

Another method of alignment is to diametrically cross the capillary tube IO from the incident laser beam. relies on the pattern of the conducted beam observed in the Referring to Figure 6, the laser beam The screen 92 is arranged perpendicular to the direction of the beam and parallel to the capillary tube lO. The pattern of not only the scattered light but also the conducted laser light after passing through the capillary It will form an image. This conducted beam is removed from the contents within the capillary tube. A vine that contains a relatively small amount of light. Figure 9 shows the relationship between a capillary and orthogonal laser beams. It represents the pattern observed when there is a match. In gel-filled tubules (The image is slightly different in oven tube mode of operation, but very similar) ), there is a bright elongated oval spot 94. This spot 94 is symmetrical about the axis of the laser beam and substantially centered about said axis . Spot 94 is located between the capillary and the separation support medium within the capillary by the conducted laser beam. It is believed to be generated by lens action based on The lens is cylindrical It essentially functions as a lens. This bright spot 94 is a far-field scattering image ( (see Figure 8 and related explanations), and the scattered image is the transmitted beam spot. It is less strong than Kit94. On both sides of the bright spot 94, there are capillary tubes. There is a symmetrical image 96.98 of the light scattered from . If the capillary tubes are not aligned 9, the resulting image will appear biased from the symmetrical image shown in Figure 9. observed. In particular, when the laser beam is moved in the positive direction of the y-axis relative to the capillary tube, the spatial distribution of the intensity of the bright spot 94 or the bright spot 94 when The end of It appears to move in the opposite direction (negative direction of the y-axis). Therefore, an appropriate host Sensors 97,99 are placed on the screen 92 to monitor the imaging action to maintain alignment. It's coming. The output from photosensor 97.99 provides feedback for automatic alignment. It can be used as a

10A to 10c show that the laser beam 29 is aligned across the capillary tube IO. It shows the evolution of the beam pattern as it moves forward. Beam 29 When focused on the center of the tube 10, the beam pattern 94 is perfectly symmetrical. , and the two photosensors 97,99 are receiving the same amount of radiation. beam 29 When translated from the center of the tubule (FIGS. 10A and 10c), spot 94 An optical imbalance is created because the brightness of the light is no longer symmetrical. Figure 1 At 0A, spot 94 is brighter at the edge closer to photosensor 99, but in Figure 10c In this case, spot 94 is brighter at the end near photosensor 97. Photosensor response difference focuses the laser beam 29 to the center of the capillary tube 10, as described below. This is a signal used for continuous tracking.

In the apparatus of FIG. 11, which schematically shows another embodiment of an automatic alignment apparatus, the apparatus shown in FIG. Instead of translating the optical fiber 28 carrying the laser beam, the laser A plano-convex lens used to focus the beam directly onto the capillary tube is translated.

Laser beam 29 from laser 26 is focused onto capillary tube 10 by plano-convex lens 120. I can match the points. Plano-convex lens 120 is supported by a positioner (not shown) . In order to focus the laser beam 29 on the center of the capillary tube 10, translational movement in the y-axis direction (or slight (lJi 94 movement) of the lens) A linear DC motor 122 (available from Foriel, Mo tormike (registered interpolation series). piezoelectric device or small Electromagnetic voice coil device suitable for distance Other linear actuators such as e-coil devices can be used This is especially mentioned. On screen 92, photosensors 97,99 detect the conducted beams. and the relative strength of the feedback signal to the motor controller 124. put out

Referring to FIG. 12, which shows one embodiment of the circuit for controller 124, a non-reversing An audio amplifier 124 is used to drive lens 120. Said circuit is balanced with the laser beam passing through the center of the tubule, as shown in Figure 10B. Ru. The imbalance in light intensity between the two photosensors is caused by the lens positioning motor 122. , focus the laser beam 29 toward the center of the capillary tube, and flatten it. To balance the point, the lens 120 is translated. the speed at which the lens is translated is proportional to the difference in light intensity (V, u, = iR, @, , w, c, /R, , , ,/)V,,,), trim pot 126.128 is circuit for bias voltage offset correction from voltage regulators 132, 134. It is provided. The trimbot 130 flattens the signal from the photosensor 97.99. to balance and eliminate inherent asymmetries between photosensors or within spots 94. It is provided.

Using the circuit described above, a mismatch of at least 365μ on the order of ±150μ It has been found that this can be corrected for a capillary with an outer diameter of 100μ and an inner diameter of 100μ.

The distance between the photo sensors 97 and 99 depends on the size of the capillary used and the capillary distance of the screen 92. , the position of the beam focus along the Z axis, and the contents of the tubule form an elliptical spot. 94 dimensions, so it is particularly important to note that will be affected.

An additional photosensor is used for the purpose of aligning the position of the laser beam in the X-axis direction. can be placed on the screen 92 spaced apart in the X-axis direction on the side of the oval spot 94. People can understand what you are saying.

Yet another example of capillary and/or laser beam alignment is described. Thin In Figure 13, which shows laser-induced fluorescence detection for tube electrophoresis, For detection by photomultiplier tube 102, reflection FA 100 of baraboloid to effectively collect and standardize the fluorescence emission from the sample within the capillary tube 10. It is used. The capillary tube is positioned to pass through the focal point 104 of the balaboloid. It's decided. Parabolo in order to prevent scattering from the capillary in the plane orthogonal to the capillary. There is a piece of material 106 placed across the mouth of the id.

For suitable criteria of fluorescence emission from the detection section of the capillary, the laser beam 108 should be directed to the focus of the paraboloid. Once, with a laser beam Once lateral alignment of the tubule with the separating tube is achieved as described above, the focus of the paraboloid The alignment of the laser beam 108 with the detection section of the capillary at The shadow of the blocking piece 106 is seen on the front surface 105 of the filter 107 that crosses the filter 110. determined by

Referring to FIGS. 14A-14c, when the laser beam is aligned with the focal point of the Polaroid, Then, the image shown in FIG. 14B is observed. The shadow of the blocking piece 106 is the background of the scattered light. Appears as two dark sections 112, 114 separated by a round . The reason for the separation in this form is not understood. The laser beam is aligned with this If the dark section 112.11 is translated upward in the X-axis direction from the 4 will be tilted downwards as shown in FIG. 14A. The laser beam is aligned When translated in the opposite direction from the original position, the dark sections 112, 114 , will be tilted upwards, as shown in Figure 14C. Therefore, the referenced beam By observing the image inside, the laser beam is narrowed at the focus of the paraboloid. One can determine that there is alignment with the detection section of the tube. Furthermore, the photomultiplier tube The alignment position of FIG. 14B can be determined by examining the signal (removing screen 105). can be determined. The signal will be at a minimum when the image of FIG. 14B is achieved. cormorant.

Alignment of the laser beam with the focus of the paraboloid is performed first, and then the laser It is understood that lateral alignment of the capillary of the beam with the separation tube can be performed.

Although the invention has been described with respect to illustrative embodiments, various modifications and improvements may be made to the invention. It will be apparent to those skilled in the art that anything can be done without departing from the scope and spirit of the invention. Dew. Therefore, the present invention should not be limited by the specific illustrated embodiments. It is limited only by the due east range of m4 dimensions.

FIG, l0B FIC;, 14A, Fl, 14B international search report , +++++n+++++v+ PCT/US 92108589,,,,, IA-++l+N+ PCT/US 92108589

Claims (1)

[Claims] a capillary tube defining a capillary channel; radiation means for irradiating the section, the radiation from the radiation means being directed to the capillary tube; The capillary tube and the tube are interconnected to create a distinct light pattern on the external screen of the tube. radiation means acting on the said capillary channel to said emitting means based on said distinct light pattern. and aligning alignment means. 2. The alignment means includes detection means for detecting the distinct light pattern. Apparatus for aligning the capillary according to item 1 with respect to a radiation source. 3. The alignment means further aligns the capillary channel with the radiation means. a control hand that controls based on feedback from the detected light pattern; 3. A device for aligning a capillary with a radiation source according to claim 2, comprising a step. 4. The radiation from the radiation means is light scattered on the inner wall of the capillary channel. the detection hand interacts with the capillary tube to produce a pattern of light; a stage arranged on the screen for detecting the pattern of scattered light; 4. A device for aligning a capillary with respect to a radiation source according to claim 3, comprising at least one sensor. Place. 5 alignment of said capillary channel to said radiating means The light scattered from two points on the inner wall of the tubular channel in the radial plane is actually considered achieved when the strength is qualitatively equal or in a given ratio detecting the light scattered from the two points and feeding it to the control means. Claim 4: There are at least two sensors located on the screen that provide a background image. Apparatus for aligning a capillary with a radiation source as described in . 6 The detection means further displays the diametrical surface of the capillary channel on the screen. 6. Apparatus for aligning a capillary with respect to a radiation source as claimed in claim 5, including means for imaging. 7 said screen is defined by a plate having an aperture, said aperture being small enough to allow light to pass through but block light scattered from the tubular channel. 7. A device for aligning a capillary tube according to claim 6 to a radiation source, the device being of small size. 8. The sensor releases the capillary of claim 7, wherein the sensor is located near the opening. A device aligned to a source. 9. The radiation means also cause fluorescence of the contents within the capillary channel. 9. A device for aligning a capillary tube according to claim 8 with a radiation source. 10 Aligning the capillary tube according to claim 9 with respect to the radiation source, wherein the radiation means comprises a laser. equipment to match. 11 said radiation from said radiation means is an elongation of radiation conducted from said radiation means; interact with the capillary tube to create a pattern of spots on the screen. and thereby the tubular channel observes the intensity distribution of the spot. 2. The radiation means according to claim 1, wherein the radiation means is considered to be aligned by device that aligns the tubules of the body with respect to the radiation source. 12 capillary tubes defining separation channels and separating the sample into its components means for causing electrophoresis of the sample; radiation means for irradiating a defined section of said capillary channel, said radiation means; The radiation from the radiation means creates a distinct light pattern on the external screen of the capillary tube. radiating means interacting with said capillary tube to emit; said capillary channel to said emitting means based on said distinct light pattern. and matching formulation means. 13 providing a capillary tube defining a capillary channel, said capillary channel; radiation means for irradiating a defined section of the capillary tube; Compatible with the capillary tube to create a distinct light pattern on the external screen of the tube. providing interacting radiation means; said capillary channel to said emitting means based on said distinct light pattern. A method of aligning a capillary to a radiation source, including aligning. 14 The step of aligning includes detecting the distinct light pattern. 14. A method of aligning a capillary to a radiation source according to claim 13. 15 The step of aligning further includes determining the detected distinct light pattern. of the capillary channel based on feedback from the radiator. Aligning the capillary of claim 14 with a radiation source, comprising controlling the alignment. Method. 16 The radiation from the radiation means is such that light scattered on the inner walls of the capillary channel interacting with the capillary tube to create a pattern from the detecting The step further includes placing the screen on the screen for detecting the scattered light. 16. The method of releasing a capillary according to claim 15, comprising providing at least one sensor with a How to align to a source. 17 The alignment of said capillary channel to said emitting means is such that said capillary channel is The light scattered from two points on the inner wall of the tubular channel in the diametrical plane is is considered to have been achieved when the strengths are substantially equal or in a given ratio. and detects the light scattered from the two points, and sends a feed to the control means. Claim: 1. There are at least two sensors located on said screen that provide feedback. 17. A method of aligning the capillary of claim 16 to a radiation source. 18 The step of detecting further includes detecting the diameter of the capillary channel. 18. The capillary of claim 17, comprising imaging a surface of the radiation source on the screen. How to align. 19 The step of detecting inhibits light scattered from the capillary channel. providing a plate having an opening dimensioned to stop the screen and defining said screen; 20. A method of aligning a capillary according to claim 18 to a radiation source. 20. The capillary of claim 19, wherein the sensor is located near the opening. How to match the radiation source to the radiation source. 21 said radiation means also cause fluorescence of the contents within said capillary channel; 21. A method of aligning a capillary according to claim 20 to a radiation source. 22. The capillary according to claim 21, wherein the radiation means comprises a laser, to a radiation source. How to align. 23. said radiation from said radiation means is an elongation of radiation conducted from said radiation means; interact with the capillary tube to create a pattern of spots on the screen. and thereby the tubular channel observes the intensity distribution of the spot. 14. The radiation means according to claim 13 is considered to be aligned with the radiation means by A method of aligning a capillary with a radiation source. 24 Prepare a capillary tube that defines a separation channel, subjecting said sample to electrophoresis for separation in minutes; radiation means for irradiating a defined section of said separation channel, comprising: Irradiation from the radiation means produces a distinct light pattern on the external screen of the capillary tube. providing radiation means for interacting with said capillary tube to produce said aligning a separation channel to the emitting means based on the distinct light pattern; An electrophoresis method comprising: